title: Expansion of adipose mesenchymal stromal cells is affected by human platelet lysate and plating density.
abstract: The composition of mesenchymal stromal cells (MSCs) changes in the course of in vitro culture expansion. Little is known how these cell preparations are influenced by culture media, plating density, or passaging. In this study, we have isolated MSCs from human adipose tissue in culture  medium supplemented with either fetal calf serum (FCS) or human platelet lysate (HPL). In addition, culture expansion was simultaneously performed at plating densities of 10 or 10,000 cells/cm(2). The use of FCS resulted in larger cells, whereas HPL significantly enhanced proliferation.  Notably, HPL also facilitated expansion for more population doublings than FCS (43 ± 3 vs. 22 ± 4 population doubling; p < 0.001), while plating density did not have a significant effect on long-term growth curves. To gain further insight into population dynamics, we  conceived a cellular automaton model to simulate expansion of MSCS. It is based on the assumptions that the number of cell divisions is limited and that due to contact inhibition proliferation occurs only at the rim of colonies. The model predicts that low plating densities result in more  heterogeneity with regard to cell division history, and favor subpopulations of higher migratory activity. In summary, HPL is a suitable serum supplement for isolation of MSC from adipose tissue and facilitates more population doublings than FCS. Cellular automaton computer simulations provided  additional insights into how complex population dynamics during long-term expansion are affected by plating density and migration.
authors.affiliation: Stem Cell Biology and Cellular Engineering, Medical Faculty, RWTH Aachen University, Aachen, Germany.
authors.forename: Dominik
authors.initials: D
authors.lastname: Cholewa
authors.forename: Thomas
authors.initials: T
authors.lastname: Stiehl
authors.forename: Anne
authors.initials: A
authors.lastname: Schellenberg
authors.forename: Gudrun
authors.initials: G
authors.lastname: Bokermann
authors.forename: Sylvia
authors.initials: S
authors.lastname: Joussen
authors.forename: Carmen
authors.initials: C
authors.lastname: Koch
authors.forename: Thomas
authors.initials: T
authors.lastname: Walenda
authors.forename: Norbert
authors.initials: N
authors.lastname: Pallua
authors.forename: Anna
authors.initials: A
authors.lastname: Marciniak-Czochra
authors.forename: Christoph V
authors.initials: CV
authors.lastname: Suschek
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Wagner
mesh_terms: D000273:Adipose Tissue / Q000166:cytology*; D001792:Blood Platelets / Q000166:cytology* / Q000378:metabolism; D002452:Cell Count; D002457:Cell Extracts / Q000494:pharmacology*; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002469:Cell Separation; D002478:Cells, Cultured; D016922:Cellular Senescence / Q000187:drug effects; D006801:Humans; D016130:Immunophenotyping; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D008954:Models, Biological; D044967:Serum; D013997:Time Factors
chemical_list: D002457:Cell Extracts